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Ultralow-input single-tube linked-read library method enables short-read second-generation sequencing systems to routinely generate highly accurate and economical long-range sequencing information

Long-range sequencing information is required for haplotype phasing, de novo assembly, and structural variation detection. Current long-read sequencing technologies can provide valuable long-range information but at a high cost with low accuracy and high DNA input requirements. We have developed a s...

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Detalles Bibliográficos
Autores principales: Chen, Zhoutao, Pham, Long, Wu, Tsai-Chin, Mo, Guoya, Xia, Yu, Chang, Peter L., Porter, Devin, Phan, Tan, Che, Huu, Tran, Hao, Bansal, Vikas, Shaffer, Justin, Belda-Ferre, Pedro, Humphrey, Greg, Knight, Rob, Pevzner, Pavel, Pham, Son, Wang, Yong, Lei, Ming
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7370886/
https://www.ncbi.nlm.nih.gov/pubmed/32540955
http://dx.doi.org/10.1101/gr.260380.119
Descripción
Sumario:Long-range sequencing information is required for haplotype phasing, de novo assembly, and structural variation detection. Current long-read sequencing technologies can provide valuable long-range information but at a high cost with low accuracy and high DNA input requirements. We have developed a single-tube Transposase Enzyme Linked Long-read Sequencing (TELL-seq) technology, which enables a low-cost, high-accuracy, and high-throughput short-read second-generation sequencer to generate over 100 kb of long-range sequencing information with as little as 0.1 ng input material. In a PCR tube, millions of clonally barcoded beads are used to uniquely barcode long DNA molecules in an open bulk reaction without dilution and compartmentation. The barcoded linked-reads are used to successfully assemble genomes ranging from microbes to human. These linked-reads also generate megabase-long phased blocks and provide a cost-effective tool for detecting structural variants in a genome, which are important to identify compound heterozygosity in recessive Mendelian diseases and discover genetic drivers and diagnostic biomarkers in cancers.